Mushroom BiologyandMushroom Products
Proceedings of the Sixth International Conference on Mushroom Biology and Mushroom Products 29th September – 3rd October, 2008 Bonn, Germany
Edited by
J.I. LelleyGAMU GmbH, Institut für Pilzforschung,Krefeld, GermanyandJ.A. BuswellInstitute of Edible Fungi, Shanghai Academyof Agricultural Sciences, Shanghai, China
Proceeding of the 6th International Conference on Mushroom Biology and Mushroom Products, 2008
Antimicrobial Activities of Four Wild Mushroom Species Collected fromTurkey
Fatih Kalyoncu and Mustafa Oskay
Celal Bayar University, Faculty of Science & Arts, Department of Biology, Muradiye,Manisa, Turkey Email: fatih.kalyoncu@bayar.edu.tr
Abstract
Four wild mushrooms, namely
Meripilus giganteus
,
Armillaria mellea, Clitocybegeotropa
and
Sparassis crispa
collected from the southwest of Turkey, were tested for theirantimicrobial activities by using the agar well diffusion method. Ethanol and chloroformextracts from the fruit bodies of these mushrooms were assayed against 10 microorganisms.The test antibiotics penicillin G, novobiocin, nalidixic acid, amphicillin, erythromycin,imipenem, chloramphenicol, vancomycin and nystatin were used for comparison. Thisresearch has shown that four wild mushrooms revealed antimicrobial activities against someGram (+) and Gram (-) bacteria, and
Candida albicans
.
Keywords:
Meripilus giganteus; Armillaria mellea; Clitocybe geotropa; Sparassis crispa;
Antimicrobial activity; Turkey
Introduction
Macrofungi have long been used as a valuable food source and as traditionalmedicines around the world, especially in Japan and China. A number of medicinalmushrooms, such as
Ganoderma lucidium
,
Tremella fuciformis
and
Lentinula edodes
, aredeemed to belong to the highest class of medicines (Wasser & Weis, 1999). Furthermore,screening programs aimed at the discovery of new bioactive metabolites from macrofungihave been performed (Rosa
et al.
2003; Dulger
et al.
2002, 2004).In research, extracts of more than 75% of the polypore mushroom species surveyed showedantimicrobial activity and 45% of 204 mushroom species inhibited the growth of a widevariety of microorganisms (Suay
et al.
2000).This experimental study is part of a program focusing on screening of wild ediblemushrooms collected from the southwest region of Turkey. The antimicrobial activities of ethanol and chloroform extracts of four wild mushrooms are reported here for the first time.
Materials and Methods
Fungi Meripilus giganteus
(Pers.) P. Karst,
Armillaria mellea
(Vahl) P. Kumm.,
Clitocybegeotropa
(Bull.) Quel. and
Sparassis
crispa
(Wulfen) Fr. were collected from the wild duringfield trips between 2006 and 2007, from the southwest of Turkey. The morphological andecological characteristics of the collected macrofungi were recorded and photographed intheir natural habitats. The specimens were identified according to macroscopic andmicroscopic features and the related literature (Watling, 1973; Moser, 1983).
Preparation of macrofungi extracts
The dried and powdered fruit bodies of macrofungi were reduced to coarse powder.Each sample (20 g) was extracted with 100 ml of ethanol and chloroform at room31
temperature, with stirring, for 4 d. The extraction solvent was evaporated to dryness. Samplesolutions were prepared by dissolving the extracts in extraction solvents.
Microbial test organisms
Test microorganisms included the following bacteria:
Staphylococcus
aureus
ATCC6538P,
Escherichia coli
,
Sarcina lutea
ATCC 9341NA,
Bacillus cereus
ATCC 7064,
Bacillussubtilis
ATCC 6633,
Salmonella
typhimurium
CCM 5445,
Proteus vulgaris
ATCC 6897
, Enterococcus faecalis
ATCC 29212,
Enterobacter cloacae
ATCC 13047D and the yeast,
Candida albicans
ATCC 10231.
Bacterial cultures were grown in Mueller-Hinton broth(Oxoid) at 37
o
C for 24 h and the yeast was incubated in glucose yeast extract broth at 30
o
Cfor 48 h. All the microorganisms were obtained from the Department of Biology, EgeUniversity, Izmir, Turkey.
Assay for antimicrobial activity
The assay was conducted as described by Perez
et al.
(1990) with slight modificationaccording to the present experimental conditions. Briefly, 50 µl inoculum (containingapproximately 10
8
bacteria per ml and 10
7
yeast per ml) were added to 25 ml melted Mueller-Hinton agar (MHA) and potato dextrose agar (PDA) medium cooled at 50 °C. This was thenpoured into 90 mm diameter sterile Petri dishes, and maintained for 1 h at room temperature.Small wells (6 mm) were cut in the agar plate using a cork borer; 60 µl of extractconcentration with a negative control (EtOH 96° and chloroform, 60 µl) were loaded in thewells. The dishes were pre-incubated at 4 °C for 2 h to allow uniform diffusion into the agar.After pre-incubation, for bacteria, the plates were incubated aerobically at 37°C for 24 h, and28 °C for 48 h for yeast. The antimicrobial activity was evaluated by measuring the inhibitionzone diameter observed. In addition, commercial antibiotics [penicillin G (10 i.u.), nalidixicacid (30 µg), novobiocin (30 µg), ampicillin (10 µg), imipenem (10 µg), erythromycin (15µg), vancomycin (30 µg), chloramphenicol (30 µg) and nystatin (10 µg)] were used aspositive control to determine the sensitivity of the strains. These studies were performed intriplicate.
Statistical Analysis
The mean values were statistically analyzed with the MINITAB Release 13.20program by the general one-way (unstacked) analysis of variance (ANOVA) to find out themost effective extracts and the most sensitive test organisms. Similarity (%) of microorganisms in relation to their susceptibility to the mushroom extracts was analyzed bythe multivariate cluster analysis according to the data obtained from well diffusion assay.
Results and Discussion
Table 1 show the antimicrobial activities of the extracts obtained from
A. mellea
,
C.geotropa
,
M. giganteus
and
S. crispa
. As clearly seen from Table 1, with an inhibition zone of 20 and 19 mm, the ethanol extracts of
A. mellea
and
C. geotropa
presented significantantibacterial activity against
B. subtilis
and
B. cereus
, respectively. The ethanol extracts of
A.mellea
and
M. giganteus
showed antiyeast activity against
C. albicans
, 19 and 20 mmrespectively. Also, the ethanol extract of
S. crispa
has antiyeast activity, with an inhibitionzone 17 mm. The ethanol extract of
A. mellea
was found to be active against
S. lutea
and
P.vulgaris
, 17 and 16 mm, respectively. Similarly, the ethanol extract of
C. geotropa
hasantibacterial activity against
P. vulgaris
(16 mm).32
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Antimicrobial Activities of Four Wild Mushroom Species Collected From Turkey
Kalyoncu F and Oskay M.
Four wild mushrooms, namely Meripilus giganteus, Armillaria mellea, Clitocybe geotropa and Sparassis crispa collected from the southwest of Turkey, were tested for their antimicrobial activities by using the agar well diffusion… (More)
Four wild mushrooms, namely Meripilus giganteus, Armillaria mellea, Clitocybe geotropa and Sparassis crispa collected from the southwest of Turkey, were tested for their antimicrobial activities by using the agar well diffusion… (More)
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